Liver-type fatty acid binding protein interacts with hepatocyte nuclear factor 4α

Hepatocyte nuclear factor 4α (HNF4α) regulates liver type fatty acid binding protein (L-FABP) gene expression. Conversely as shown herein, L-FABP structurally and functionally also interacts with HNF4α. Fluorescence resonance energy transfer (FRET) between Cy3-HNF4α (donor) and Cy5-L-FABP (acceptor) as well as FRET microscopy detected L-FABP in close proximity (∼80 Å) to HNF4α, binding with high affinity K_d ∼250–300 nM. Circular dichroism (CD) determined that the HNF4α/L-FABP interaction altered protein secondary structure. Finally, L-FABP potentiated transactivation of HNF4α in COS7 cells. Taken together, these data suggest that L-FABP provides a signaling path to HNF4α activation in the nucleus.     

Fibrillar seeds alleviate amyloid-β cytotoxicity by omitting formation of higher-molecular-weight oligomers

Amyloid-β (Aβ) peptides can exist in distinct forms including monomers, oligomers and fibrils, consisting of increased numbers of monomeric units. Among these, Aβ oligomers are implicated as the primary toxic species as pointed by multiple lines of evidence. It has been suggested that toxicity could be rendered by the soluble higher-molecular-weight (high-n) Aβ oligomers. Yet, the most culpable form in the pathogenesis of Alzheimer’s disease (AD) remains elusive. Moreover, the potential interaction among the insoluble fibrils that have been excluded from the responsible aggregates in AD development, Aβ monomers and high-n oligomers is undetermined. Here, we report that insoluble Aβ fibrillar seeds can interact with Aβ monomers at the stoichiometry of 1:2 (namely, each Aβ molecule of seed can bind to two Aβ monomers at a time) facilitating the fibrillization by omitting the otherwise mandatory formation of the toxic high-n oligomers during the fibril maturation. As a result, the addition of exogenous Aβ fibrillar seeds is seen to rescue neuronal cells from Aβ cytotoxicity presumably exerted by high-n oligomers, suggesting an unexpected protective role of Aβ fibrillar seeds.     

Flagella proteins contribute to the production of outer membrane vesicles from Escherichia coli W3110

Gram-negative bacteria, including Escherichia coli, release outer membrane vesicles (OMVs) that are derived from the bacterial outer membrane. OMVs contribute to bacterial cell–cell communications and host–microbe interactions by delivering components to locations outside the bacterial cell. In order to explore the molecular machinery involved in OMV biogenesis, the role of a major OMV protein was examined in the production of OMVs from E. coli W3110, which is a widely used standard E. coli K-12 strain. In addition to OmpC and OmpA, which are used as marker proteins for OMVs, an analysis of E. coli W3110 OMVs revealed that they also contain abundant levels of FliC, which is also known as flagellin. A membrane-impermeable biotin-labeling reagent did not label FliC in intact OMVs, but labeled FliC in sonically disrupted OMVs, suggesting that FliC is localized in the lumen of OMV. Compared to the parental strain expressing wild-type fliC, an E. coli strain with a fliC-null mutation produced reduced amounts of OMVs based on both protein and phosphate levels. In addition, an E. coli W3110-derived strain with a null-mutation in flgK, which encodes flagellar hook-associated protein that is essential along with FliC for flagella synthesis, also produced fewer OMVs than the parental strain. Taken together, these results indicate that the ability to form flagella, including the synthesis of flagella proteins, affects the production of E. coli W3110 OMVs.     

Amyloplast displacement is necessary for gravisensing in Arabidopsis shoots as revealed by a centrifuge microscope

The starch‐statolith hypothesis proposes that starch‐filled amyloplasts act as statoliths in plant gravisensing, moving in response to the gravity vector and signaling its direction. However, recent studies suggest that amyloplasts show continuous, complex movements in Arabidopsis shoots, contradicting the idea of a so‐called ‘static’ or ‘settled’ statolith. Here, we show that amyloplast movement underlies shoot gravisensing by using a custom‐designed centrifuge microscope in combination with analysis of gravitropic mutants. The centrifuge microscope revealed that sedimentary movements of amyloplasts under hypergravity conditions are linearly correlated with gravitropic curvature in wild‐type stems. We next analyzed the hypergravity response in the shoot gravitropism 2 (sgr2) mutant, which exhibits neither a shoot gravitropic response nor amyloplast sedimentation at 1  g . sgr2 mutants were able to sense and respond to gravity under 30  g conditions, during which the amyloplasts sedimented. These findings are consistent with amyloplast redistribution resulting from gravity‐driven movements triggering shoot gravisensing. To further support this idea, we examined two additional gravitropic mutants, phosphoglucomutase (pgm) and sgr9, which show abnormal amyloplast distribution and reduced gravitropism at 1  g . We found that the correlation between hypergravity‐induced amyloplast sedimentation and gravitropic curvature of these mutants was identical to that of wild‐type plants. These observations suggest that Arabidopsis shoots have a gravisensing mechanism that linearly converts the number of amyloplasts that settle to the ‘bottom’ of the cell into gravitropic signals. Further, the restoration of the gravitropic response by hypergravity in the gravitropic mutants that we tested indicates that these lines probably have a functional gravisensing mechanism that is not triggered at 1  g .     

Vertical migration patterns in phytoplankton of mixed species composition

The vertical distribution and migration of phytoplankton composed of seven organisms in a small eutrophic pool is described. Vertical migration during a 24 h period is shown to be exhibited by Pandorina morum, Mallomonas tonsurata, Chroomonas pusilla (Rhodomonas) and Ochromonas sp. in the epilimnion and Cryptomonas rufescens in the surface of the hypolimnion. The degree of horizontal variation in distribution of the organisms is discussed and found to be greatest for the microflagellates. Changes in vertical distribution are shown to be significant and result from active movement of the cells rather than from either water movement or the combined variation associated with patchiness in distribution and counting errors.

Differences between the behaviour patterns of the different algae are evident and may be used to explain Hutchinson's “paradox of the plankton”.     

Using laser scattering to identify diatoms and conduct aggregation experiments

Laser in situ scattering and transmissometry (LISST) instruments are used to measure particle size distributions (PSDs) and volume concentrations in water. For populations of regularly shaped non-spherical particles, such as phytoplankton, the PSD produces a ‘scattering signature’ that corresponds to the shape of the particles. The objectives of this research were to describe the scattering signatures of six diatom species and to determine whether LISST instruments can be used as a tool to measure the aggregation of diatoms into larger particles. The scattering signatures of Chaetoceros muelleri var. subsalsum, Coscinodiscus wailesii, Thalassiosira weissflogii, Phaeodactylum tricornutum, Skeletonema costatum and S. marinoi were measured. The scattering signatures of individual species were consistent over time in batch culture and there were clear differences between species in terms of peak location, peak width, and relative peak height in the PSD. LISST was used to non-destructively follow the formation of diatom aggregates in the laboratory. Both rolling and warming cultures of S. costatum caused the cell chains to form aggregates, resulting in a change in the PSD, with a shift in peak position towards larger size bins. These experiments showed that the scattering signatures of unaggregated diatom species are conservative and that LISST instruments are useful tools to investigate the factors affecting diatom aggregation and disaggregation, with potential applications both in the laboratory and field.     

小哺乳动物化石牙齿釉质碳、氧同位素测试方法简介及应用前景

探讨古环境和古气候变化与哺乳动物演化之间的关系是目前古生物学研究领域中的一个热点,而哺乳动物化石牙齿釉质的碳、氧同位素分析是恢复古环境和古气候的一个重要手段。以往的哺乳动物化石牙齿釉质稳定同位素分析多集中在大哺乳动物化石,这主要是受到技术手段的限制,所需的样品量较大所决定的。但最近几年随着激光和离子显微探针技术的应用,对小哺乳动物化石(如啮齿类和兔形类)的牙齿釉质碳、氧同位素的分析和应用日趋成熟和广泛。除了传统的化学处理方法之外,对小哺乳动物化石牙齿釉质碳、氧同位素的分析还有以下三种方法:1)激光剥蚀气相色谱/同位素比值质谱分析;2)直接激光氟化技术;3)离子显微探针技术(SHRIMPII)。这些技术需要的样品量少,对标本的破损小,准确度和精密度高,所以在小哺乳动物化石和一些珍贵标本(如古人类化石)的稳定同位素分析中起到了重要作用。相对于大哺乳动物化石,小哺乳动物化石数量多、演化速度快,更能反映多个层位长时间序列的古环境和气候变化;而且小哺乳动物通常没有长距离迁徙的行为,栖息地局限,所以更能准确反映化石埋藏地点的古环境和气候状况。     

小金海棠果实表皮制片方法的比较研究

为了简便而快速制作适用于光学显微镜观测的苹果属植物果实表皮标本,采用7种方法分别对小金海棠果实表皮进行整体制片后于光学显微镜下观察其形态特征,比较制片效果。结果表明:用离析刮片法、指甲油印迹法、刮片法和煮沸剥离刮片法制备的标本在光学显微镜下均呈现出清晰结构,其中煮沸剥离刮片法的制片效果最佳。因此,煮沸剥离刮片法最适用于苹果属植物果实表皮整体制片,该方法操作简便且效率高。     

He-Ne激光和增强UV-B辐射对小麦幼叶叶绿素荧光和Rubisco活化酶的影响

选用小麦‘ML7113’品种为材料,人工模拟He-Ne激光(5mJ·s-1·mm-2)、增强UV-B(10.8kJ·m-2·d-1)辐射及两者复合辐照进行处理,利用叶绿素荧光仪、考马斯亮蓝G-250染色法和PCR技术研究7d龄小麦幼苗叶绿素荧光特性、Rubisco活化酶含量、基因表达量及其基因序列的变化。结果表明:(1)与对照组相比,增强UV-B辐射后,小麦幼苗叶绿素荧光特性减弱,Rubisco活化酶含量及其基因表达量均下降;而低剂量的He-Ne激光辐照后能够在一定程度上修复经UV-B辐射后对小麦幼苗叶绿素荧光特性所造成的损伤,且使Rubisco活化酶含量及其基因表达量上升。(2)与对照组相比,经He-Ne激光和增强UV-B辐射以及两者复合辐照处理后基因序列均出现两个相同的点突变,但并未造成氨基酸序列的变化。研究认为,低剂量He-Ne激光辐照能够在一定程度上修复受UV-B辐射小麦幼苗叶绿素荧光活性、Rubisco活化酶含量及其基因表达量的降低;He-Ne激光和增强UV-B辐射对小麦幼苗Rubisco活化酶活性的影响可能发生在其转录水平,从而使小麦光合能力发生相应的变化。     

He-Ne激光预处理对镉胁迫下小麦幼苗生理特性的影响

用He-Ne激光（波长632.8 nm,辐射剂量5.43 mW/mm²）对萌动小麦种子辐照5 min,待幼苗长至一叶一心时,用150 μmol/L CdCl₂溶液进行胁迫处理,研究He-Ne激光预处理对镉（Cd²⁺）胁迫下小麦幼苗生长发育和生理特性的影响。结果显示:He-Ne激光预处理能显著降低Cd²⁺胁迫下小麦幼苗中丙二醛（MDA）、过氧化氢（H₂O₂）含量及超氧自由基（O^-·₂）产生速率,显著提高幼苗叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)、抗坏血酸氧化酶（APX）活性,并使叶片抗氧化物质谷胱甘肽(GSH)和抗坏血酸(AsA)含量以及幼苗株高、根长和干重增加。研究表明,He-Ne激光预处理可有效缓解镉胁迫对小麦幼苗生长的抑制作用,并通过促进其幼苗中酶类和非酶类抗氧化剂的产生,有效减少镉胁迫产生的脂质过氧化物含量,从而提高其耐镉性。